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Wip1 phosphatase is associated with chromatin and dephosphorylates γH2AX to promote checkpoint inhibition

Oncogene volume 29, pages 2281–2291 (2010)Cite this article

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Abstract

DNA double-stranded breaks (DSBs) elicit a checkpoint response that causes a delay in cell cycle progression. Early in the checkpoint response, histone H2AX is phosphorylated in the chromatin region flanking the DSB by ATM/ATR and DNA-PK kinases. The resulting foci of phosphorylated H2AX (γ-H2AX) serve as a platform for recruitment and retention of additional components of the checkpoint-signaling cascade that enhance checkpoint signaling and DSB repair. Upon repair, both the assembled protein complexes and the chromatin modifications are removed to quench the checkpoint signal. In this study, we show that the DNA damage-responsive Wip1 phosphatase is bound to chromatin. Moreover, Wip1 directly dephosphorylates γ-H2AX and cells depleted of Wip1 fail to dephosphorylate γ-H2AX during checkpoint recovery. Conversely, premature activation of Wip1 leads to displacement of MDC1 from damage foci and prevents activation of the checkpoint. Taken together, our data show that Wip1 has an essential role in dephosphorylation of γ-H2AX to silence the checkpoint and restore chromatin structure once DNA damage is repaired.

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Acknowledgements

We are thankful to Dr Lukas for providing MDC–EGFP plasmid, Dr Donehower for Wip1–FLAG, Dr Dansen and P Charitou for providing reagents, Livio Kleij for maintenance of the microscopes and Monica Alvarez for critical reading of the article. RHM and LM were funded by the Netherlands Genomics Initiative of NWO and the Dutch Cancer Society (UU2006-3579), LM by Grant Agency of the Czech Republic (P305/10/P420), AL by the Wenner-Gren foundation and by the Netherlands Organization for Scientific Research (NWO) (ZonMw 916.86.083) and HV by the Center for Translational Molecular Medicine (CTMM).

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  1. Department of Medical Oncology and Cancer Genomics Center, University Medical Center Utrecht, Utrecht, The Netherlands

    L Macůrek, A Lindqvist, O Voets, J Kool & R H Medema

  2. Department of Genome Integrity, Institute of Molecular Genetics, v.v.i., Academy of Sciences of the Czech Republic, Prague, Czech Republic

    L Macůrek

  3. Department of Physiological Chemistry and Cancer Genomics Center, University Medical Center Utrecht, Utrecht, The Netherlands

    H R Vos

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  1. L Macůrek
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Correspondence to R H Medema.

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Macůrek, L., Lindqvist, A., Voets, O. et al. Wip1 phosphatase is associated with chromatin and dephosphorylates γH2AX to promote checkpoint inhibition. Oncogene 29, 2281–2291 (2010). https://doi.org/10.1038/onc.2009.501

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